A lactoferrin-derived peptide LFDP1 alleviates experimental NEC via blocking ACSL4-LPCAT3 axis

Abstract

This study investigated the anti-necrotizing enterocolitis (NEC) effects of LFDP1, a lactoferrin-derived peptide previously identified as up-regulated in human milk from mothers of preterm infants. Using both in vivo and in vitro models, we evaluated LFDP1's potential to attenuate NEC development. In NEC rat pups, LFDP1 administration significantly reduced the severity of clinical symptoms and pathological features. At the cellular level, LFDP1 restored viability and enhanced migration and proliferation of IEC6 and FHC cells. Molecular analysis revealed that LFDP1 downregulated key NEC process indicators (TLR4, ACSL4, and LPCAT3) while upregulating protective factors (ZO-1 and GPX4). RNA sequencing of LFDP1-treated FHC cells identified 117 differentially expressed mRNAs, with pathway analysis highlighting the involvement of polyunsaturated fatty acid biosynthesis and fatty acid metabolism. Further investigation showed that LFDP1 suppressed ferroptosis by inhibiting the ACSL4-LPCAT3 signaling axis, a finding corroborated in human NEC intestinal samples. Notably, LFDP1 treatment reduced oxidative stress and modulated the expression of ferroptosis-related proteins in vitro. These findings suggest that LFDP1 alleviates experimental NEC by repressing polyunsaturated fatty acid production and inhibiting ferroptosis via the ACSL4-LPCAT3 pathway. Our study not only elucidates a potential mechanism for LFDP1's protective effects but also contributes to the broader understanding of how breast milk composition may protect newborns from diseases like NEC.

Keywords

• Human milk
• Peptide
• Neonate
• Necrotizing enterocolitis
• Ferroptosis